免疫复合物解离剂的研究及在HCV抗原检测中的应用效果分析  

Investigation of immune complex dissociation reagents and their application in detecting HCV antigen

作　　者：杨蓉[1] 龙润乡[1] 李华[1] 谭振国[1] 白惠珠[1] 杨婷[1] 蒋蕊鞠[1] 谢忠平[1] 

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所病原生物技术诊断研究组,昆明650118

出　　处：《免疫学杂志》2012年第7期620-623,共4页Immunological Journal

基　　金：国家863项目(2007AA02Z480);云南省联合支持国家计划项目(2008GA008)

摘　　要：目的探索一种可以使免疫复合物发生解离的方法,并进行应用效果分析。方法通过对HAV抗原抗体复合物的解离,进行酸化剂、去垢剂的组合及解离时间、温度等相关条件筛选,优化出最佳的解离方法,应用于1 370份HCV血清样品,对解离前后的抗原阳性率进行统计分析。结果 160 EU/ml的HAV抗原能被效价为20 U/ml的HAV抗血清完全中和;只单独使用去垢剂对免疫复合物无解离效果;0.1 mol/LGly-HCL能解离抗原抗体复合物,加入1种或几种去垢剂后则其解离效果有不同程度的提高,其中以Gly-HCl+7%TritonX-100+5%CHAP的效果最好(q=27.177,P<0.01);免疫复合物在加入解离剂后在室温60 min、37℃30 min、60℃10 min为本试验较好的解离条件,其中以37℃30 min效果最佳;应用Gly-HCl+5%CHAPS+7%TritonX-100在37℃对样品进行预处理30 min后,可使HCV抗原阳性检出率由解离前的33.3%提升至75.56%,解离前后之间的阳性率差异具有显著地统计学意义(χ2=32.34,P<0.01)。结论酸化剂能将HAV及HCV抗原抗体免疫复合物解离,添加去垢剂后,解离效果好于单独使用酸化剂,且联合添加好于单独添加。Antigen is one of infectious markers prior to antibody. Antigen test has been proven to be able to remedy loss of detection in window period caused by antibody test. Currently, hat antigens forming immune complex with antibodies leads to lower detection rate is exactly the reason why antigen test is less used. The optimal method of dissociation was optimized by screening time ＆ temperature arid etc. regarding HAV antigen-antibody complex, time of acidizing and composition of detergents, and was applied in 1370 HCV serum samples for making statistics and analysis of antigen positive rate before and after dissociation. We inferred from results the optimal condition of dissociation was presented by using Gly-HC1 ＋ 7%TritonX-100 ＋ 5% CHAP for 30 minutes at 37℃. The application of this method in HCV antigen test has been proven to enable HCV antigen positive detection rate to improve to 75.56% from 33.3% before dissociation, and to enable differences in positive rate before and after dissociation to have markedly statistical significances （X2 = 32.34, P 〈 0.01）. Hence, it can be considered that this method can dissociate immune complex formed by HCV so as to improve detection rate of HCV antigen.

关 键 词：甲型肝炎病毒 丙型肝炎病毒 抗原抗体复合物 解离 抗原检测 

分 类 号：R392-33[医药卫生—免疫学]